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Waters Corporation seppak c18 cartridges
Seppak C18 Cartridges, supplied by Waters Corporation, used in various techniques. Bioz Stars score: 94/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/seppak c18 cartridges/product/Waters Corporation
Average 94 stars, based on 7 article reviews
Price from $9.99 to $1999.99
seppak c18 cartridges - by Bioz Stars, 2020-04
94/100 stars

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Centrifugation:

Article Title: A conserved major facilitator superfamily member orchestrates a subset of O-glycosylation to aid macrophage tissue invasion
Article Snippet: The digests were acidified with 12 µL trifluoroacetic acid (TFA), 37°C, 20 min and cleared by centrifugation at 10,000 g, 10 min. .. The cleared acidified digests were loaded onto equilibrated SepPak C18 cartridges (Waters) followed by 3 × CV 0.1% TFA wash. Digests were labeled on the column by adding 5 mL 30 mM NaBH3 CN and 0.2% formaldehyde (COH2 ) in 50 mM sodium phosphate buffer pH 7.5 (Light, mrva3102 ), or 30 mM NaBH3 CN and 0.2% deuterated formaldehyde (COD2 ) in 50 mM sodium phosphate buffer pH 7.5 (Medium, control).

Article Title: Dynamic Impacts of the Inhibition of the Molecular Chaperone Hsp90 on the T-Cell Proteome Have Implications for Anti-Cancer Therapy
Article Snippet: After centrifugation and protein concentration measurements, equimolar extracts from light/heavy- or heavy/medium-labelled cells were combined, alkylated with iodoacetamide and digested as described . .. The obtained peptide mixtures (200 µg total material) were desalted on SepPak C18 cartridges (Waters Corp., Milford, MA), dried, dissolved in 4M Urea with 0.1% Ampholytes pH 3–10 (GE Healthcare) and fractionated by off-gel focusing as described .

Article Title: De novo activating mutations drive clonal evolution and enhance clonal fitness in KMT2A-rearranged leukemia
Article Snippet: .. Following centrifugation, supernatants were collected and samples were desalted with SepPak C18 cartridges (Waters) by washing the columns with acetonitrile, conditioning and loading samples in 0.1% (v/v) trifluoroacetic acid, washing with 0.1% formic acid and eluting peptides with 80% (v/v) acetonitrile/0.1% (v/v) formic acid. .. For the control mix, four KMT2A-MLLT3 + Empty-GFP samples (all labeled with TMT 126) were mixed at this stage and split into three equal volumes.

Construct:

Article Title: Phosphoproteome Profiling Reveals Circadian Clock Regulation of Posttranslational Modifications in the Murine Hippocampus
Article Snippet: For proteomic analysis, 0.1 mg of resulting peptides were fractionated through an in-house constructed strong cation exchange (SCX) column with five pH fractions (pH 4.0, 6.0, 8.0, 10.0, and 12.0) followed by desalting with in-house C18 desalting cartridges and dried in a speed-vac prior to LC-MS analysis. .. The remaining 1.1 mg of tryptic peptides were desalted by SepPak C18 cartridges (Waters, Mississauga, ON, Canada), dried, and SCX fractionated into four fractions (pH 4.0, 6.0, 8.0, and 12.0) prior to phosphoproteome enrichment.

Incubation:

Article Title: Phosphoproteome Profiling Reveals Circadian Clock Regulation of Posttranslational Modifications in the Murine Hippocampus
Article Snippet: Hippocampal lysates (1 mg) and super SILAC-labeled cell lysates (0.2 mg from each of Neuro-2a, AtT-20, mHypoE-N38, CLU-181, and CLU-188 cells) were mixed at a 1:1 weight ratio, and SDS in solution was removed by an overnight incubation at −20°C in five volumes of ice-cold precipitation buffer [acetone/ethanol/acetic acid (v/v/v) = 50/50/0.1]. .. The remaining 1.1 mg of tryptic peptides were desalted by SepPak C18 cartridges (Waters, Mississauga, ON, Canada), dried, and SCX fractionated into four fractions (pH 4.0, 6.0, 8.0, and 12.0) prior to phosphoproteome enrichment.

Article Title: De novo activating mutations drive clonal evolution and enhance clonal fitness in KMT2A-rearranged leukemia
Article Snippet: The reaction was quenched by adding 8 µl of 5% hydroxylamine and samples were incubated at RT for 15 min. RapiGest was then precipitated by addition of 10 µl 10% trifluoroacetic acid and incubation at 37 °C for 45 min. .. Following centrifugation, supernatants were collected and samples were desalted with SepPak C18 cartridges (Waters) by washing the columns with acetonitrile, conditioning and loading samples in 0.1% (v/v) trifluoroacetic acid, washing with 0.1% formic acid and eluting peptides with 80% (v/v) acetonitrile/0.1% (v/v) formic acid.

Mass Spectrometry:

Article Title: Phosphoproteome Profiling Reveals Circadian Clock Regulation of Posttranslational Modifications in the Murine Hippocampus
Article Snippet: Paragraph title: Proteomic Analysis of Hippocampal Tissues Using Super-SILAC-Based Quantitative MS ... The remaining 1.1 mg of tryptic peptides were desalted by SepPak C18 cartridges (Waters, Mississauga, ON, Canada), dried, and SCX fractionated into four fractions (pH 4.0, 6.0, 8.0, and 12.0) prior to phosphoproteome enrichment.

Article Title: Quantifying exosome secretion from single cells reveals a modulatory role for GPCR signaling
Article Snippet: In brief, cells were lysed in denaturing lysis buffer supplemented with phosphatase inhibitors, lysates were digested overnight with sequence grade–modified trypsin (Promega), and 0.5-mg aliquots were desalted on SepPak-C18 cartridges (Waters). .. Then, phosphopeptides were captured through metal oxide affinity chromatography with in-house–prepared TiOx STAGE tips (packed with 2.5 mg 10-µm TiO2 beads; GL Sciences) in the presence of 300 mg/ml lactic acid to suppress background binding of acidic peptides, eluted with piperidine, desalted with in-house–prepared STAGE tips packed with SDB-XC (3M Empore), dried, and redissolved, and then 50% was injected into a nano–liquid chromatography–tandem mass spectrometry system (Ultimate 3000 nanoLC coupled on-line to a QExactive mass spectrometer; Thermo Fisher Scientific) operated as described previously by .

Article Title: Dynamic Impacts of the Inhibition of the Molecular Chaperone Hsp90 on the T-Cell Proteome Have Implications for Anti-Cancer Therapy
Article Snippet: The obtained peptide mixtures (200 µg total material) were desalted on SepPak C18 cartridges (Waters Corp., Milford, MA), dried, dissolved in 4M Urea with 0.1% Ampholytes pH 3–10 (GE Healthcare) and fractionated by off-gel focusing as described . .. Samples were analysed on a hybrid linear trap LTQ-Orbitrap Velos mass spectrometer (Thermo Fisher, Bremen, Germany) interfaced via a nanospray source to a Dionex RSLC 3000 nanoHPLC system (Dionex, Sunnyvale, CA, USA).

Article Title: Targeting LIF-mediated paracrine interaction for pancreatic cancer therapy and monitoring
Article Snippet: Digested peptides were de-salted using SepPak C18 cartridges (Waters) and fractionated into 28 fractions by using strong-cation exchange (SCX) chromatography (PolySULFOETHYL Aspartamide, 2.1 mm × 200 mm, 5 μm, PolyLC) with 70 min gradient at flow rate of 0.3 mL/min as reported previously . .. Peptides in each fraction were then desalting by homemade C18 StageTip for MS analysis.

Article Title: De novo activating mutations drive clonal evolution and enhance clonal fitness in KMT2A-rearranged leukemia
Article Snippet: Cysteine disulfide bonds were reduced with 5 mM DTT for 30 min at 56 °C, alkylated with 10 mM iodoacetamide for 30 min at room temperature in the dark, and proteins were digested with Mass Spectrometry Grade Lysyl Endopeptidase (Wako, Osaka, Japan) at enzyme:protein ratio 1:50, at 37 °C overnight. .. Following centrifugation, supernatants were collected and samples were desalted with SepPak C18 cartridges (Waters) by washing the columns with acetonitrile, conditioning and loading samples in 0.1% (v/v) trifluoroacetic acid, washing with 0.1% formic acid and eluting peptides with 80% (v/v) acetonitrile/0.1% (v/v) formic acid.

High Performance Liquid Chromatography:

Article Title: The Nicotiana tabacum ABC transporter NtPDR3 secretes O-methylated coumarins in response to iron deficiency
Article Snippet: Metabolites were then retained on SepPak C18 cartridges (Waters), eluted with 2 ml of methanol, and stored at –80 °C until further analysis. .. HPLC/DAD-ESI-MS (time-of flight, TOF) and HPLC/ESI-MS/MS (ion trap) analyses were carried out as described by ).

Flow Cytometry:

Article Title: Targeting LIF-mediated paracrine interaction for pancreatic cancer therapy and monitoring
Article Snippet: .. Digested peptides were de-salted using SepPak C18 cartridges (Waters) and fractionated into 28 fractions by using strong-cation exchange (SCX) chromatography (PolySULFOETHYL Aspartamide, 2.1 mm × 200 mm, 5 μm, PolyLC) with 70 min gradient at flow rate of 0.3 mL/min as reported previously . .. Peptides in each fraction were then desalting by homemade C18 StageTip for MS analysis.

RIA Assay:

Article Title: Temperature has a major influence on cardiac natriuretic peptide in salmon
Article Snippet: .. The plasma samples were extracted with SepPak C18 cartridges (Waters, Milford, MA, USA) as described previously ( ) for use in sCP radioimmunoassay. .. Aliquots of the homogenates obtained during RNA purification (see above) were used to measure immunoreactive sCP (ir-sCP) in the tissue samples.

Chromatography:

Article Title: Quantifying exosome secretion from single cells reveals a modulatory role for GPCR signaling
Article Snippet: In brief, cells were lysed in denaturing lysis buffer supplemented with phosphatase inhibitors, lysates were digested overnight with sequence grade–modified trypsin (Promega), and 0.5-mg aliquots were desalted on SepPak-C18 cartridges (Waters). .. Then, phosphopeptides were captured through metal oxide affinity chromatography with in-house–prepared TiOx STAGE tips (packed with 2.5 mg 10-µm TiO2 beads; GL Sciences) in the presence of 300 mg/ml lactic acid to suppress background binding of acidic peptides, eluted with piperidine, desalted with in-house–prepared STAGE tips packed with SDB-XC (3M Empore), dried, and redissolved, and then 50% was injected into a nano–liquid chromatography–tandem mass spectrometry system (Ultimate 3000 nanoLC coupled on-line to a QExactive mass spectrometer; Thermo Fisher Scientific) operated as described previously by .

Article Title: Targeting LIF-mediated paracrine interaction for pancreatic cancer therapy and monitoring
Article Snippet: .. Digested peptides were de-salted using SepPak C18 cartridges (Waters) and fractionated into 28 fractions by using strong-cation exchange (SCX) chromatography (PolySULFOETHYL Aspartamide, 2.1 mm × 200 mm, 5 μm, PolyLC) with 70 min gradient at flow rate of 0.3 mL/min as reported previously . .. Peptides in each fraction were then desalting by homemade C18 StageTip for MS analysis.

Cell Culture:

Article Title: Targeting LIF-mediated paracrine interaction for pancreatic cancer therapy and monitoring
Article Snippet: For CM stimulation assays on cultured cells, CM were used freshly or stored at 4°C for no more than 3 days. .. Digested peptides were de-salted using SepPak C18 cartridges (Waters) and fractionated into 28 fractions by using strong-cation exchange (SCX) chromatography (PolySULFOETHYL Aspartamide, 2.1 mm × 200 mm, 5 μm, PolyLC) with 70 min gradient at flow rate of 0.3 mL/min as reported previously .

Protein Concentration:

Article Title: Dynamic Impacts of the Inhibition of the Molecular Chaperone Hsp90 on the T-Cell Proteome Have Implications for Anti-Cancer Therapy
Article Snippet: After centrifugation and protein concentration measurements, equimolar extracts from light/heavy- or heavy/medium-labelled cells were combined, alkylated with iodoacetamide and digested as described . .. The obtained peptide mixtures (200 µg total material) were desalted on SepPak C18 cartridges (Waters Corp., Milford, MA), dried, dissolved in 4M Urea with 0.1% Ampholytes pH 3–10 (GE Healthcare) and fractionated by off-gel focusing as described .

Sequencing:

Article Title: Quantifying exosome secretion from single cells reveals a modulatory role for GPCR signaling
Article Snippet: .. In brief, cells were lysed in denaturing lysis buffer supplemented with phosphatase inhibitors, lysates were digested overnight with sequence grade–modified trypsin (Promega), and 0.5-mg aliquots were desalted on SepPak-C18 cartridges (Waters). .. Then, phosphopeptides were captured through metal oxide affinity chromatography with in-house–prepared TiOx STAGE tips (packed with 2.5 mg 10-µm TiO2 beads; GL Sciences) in the presence of 300 mg/ml lactic acid to suppress background binding of acidic peptides, eluted with piperidine, desalted with in-house–prepared STAGE tips packed with SDB-XC (3M Empore), dried, and redissolved, and then 50% was injected into a nano–liquid chromatography–tandem mass spectrometry system (Ultimate 3000 nanoLC coupled on-line to a QExactive mass spectrometer; Thermo Fisher Scientific) operated as described previously by .

Article Title: Targeting LIF-mediated paracrine interaction for pancreatic cancer therapy and monitoring
Article Snippet: The concentrated proteins were dissolved in 8 M urea buffer containing 20 mM HEPES (pH 8), then reduced by dithiothreitol (Sigma), alkylated by iodoacetamide (Sigma), and digested by sequencing-grade trypsin (Promega). .. Digested peptides were de-salted using SepPak C18 cartridges (Waters) and fractionated into 28 fractions by using strong-cation exchange (SCX) chromatography (PolySULFOETHYL Aspartamide, 2.1 mm × 200 mm, 5 μm, PolyLC) with 70 min gradient at flow rate of 0.3 mL/min as reported previously .

Sonication:

Article Title: A conserved major facilitator superfamily member orchestrates a subset of O-glycosylation to aid macrophage tissue invasion
Article Snippet: The lysed material was left on ice for 40 min with occasional vortexing followed by probe sonication (5 s sonication, 5 s pause, 6 cycles at 60% amplitude). .. The cleared acidified digests were loaded onto equilibrated SepPak C18 cartridges (Waters) followed by 3 × CV 0.1% TFA wash. Digests were labeled on the column by adding 5 mL 30 mM NaBH3 CN and 0.2% formaldehyde (COH2 ) in 50 mM sodium phosphate buffer pH 7.5 (Light, mrva3102 ), or 30 mM NaBH3 CN and 0.2% deuterated formaldehyde (COD2 ) in 50 mM sodium phosphate buffer pH 7.5 (Medium, control).

Injection:

Article Title: Quantifying exosome secretion from single cells reveals a modulatory role for GPCR signaling
Article Snippet: In brief, cells were lysed in denaturing lysis buffer supplemented with phosphatase inhibitors, lysates were digested overnight with sequence grade–modified trypsin (Promega), and 0.5-mg aliquots were desalted on SepPak-C18 cartridges (Waters). .. Then, phosphopeptides were captured through metal oxide affinity chromatography with in-house–prepared TiOx STAGE tips (packed with 2.5 mg 10-µm TiO2 beads; GL Sciences) in the presence of 300 mg/ml lactic acid to suppress background binding of acidic peptides, eluted with piperidine, desalted with in-house–prepared STAGE tips packed with SDB-XC (3M Empore), dried, and redissolved, and then 50% was injected into a nano–liquid chromatography–tandem mass spectrometry system (Ultimate 3000 nanoLC coupled on-line to a QExactive mass spectrometer; Thermo Fisher Scientific) operated as described previously by .

Binding Assay:

Article Title: Quantifying exosome secretion from single cells reveals a modulatory role for GPCR signaling
Article Snippet: In brief, cells were lysed in denaturing lysis buffer supplemented with phosphatase inhibitors, lysates were digested overnight with sequence grade–modified trypsin (Promega), and 0.5-mg aliquots were desalted on SepPak-C18 cartridges (Waters). .. Then, phosphopeptides were captured through metal oxide affinity chromatography with in-house–prepared TiOx STAGE tips (packed with 2.5 mg 10-µm TiO2 beads; GL Sciences) in the presence of 300 mg/ml lactic acid to suppress background binding of acidic peptides, eluted with piperidine, desalted with in-house–prepared STAGE tips packed with SDB-XC (3M Empore), dried, and redissolved, and then 50% was injected into a nano–liquid chromatography–tandem mass spectrometry system (Ultimate 3000 nanoLC coupled on-line to a QExactive mass spectrometer; Thermo Fisher Scientific) operated as described previously by .

Molecular Weight:

Article Title: The Nicotiana tabacum ABC transporter NtPDR3 secretes O-methylated coumarins in response to iron deficiency
Article Snippet: Metabolites were then retained on SepPak C18 cartridges (Waters), eluted with 2 ml of methanol, and stored at –80 °C until further analysis. .. Molecular formulae were assigned based on the exact molecular weight with errors < 5 ppm.

Labeling:

Article Title: A conserved major facilitator superfamily member orchestrates a subset of O-glycosylation to aid macrophage tissue invasion
Article Snippet: .. The cleared acidified digests were loaded onto equilibrated SepPak C18 cartridges (Waters) followed by 3 × CV 0.1% TFA wash. Digests were labeled on the column by adding 5 mL 30 mM NaBH3 CN and 0.2% formaldehyde (COH2 ) in 50 mM sodium phosphate buffer pH 7.5 (Light, mrva3102 ), or 30 mM NaBH3 CN and 0.2% deuterated formaldehyde (COD2 ) in 50 mM sodium phosphate buffer pH 7.5 (Medium, control). ..

Article Title: Deconstruction of O‐glycosylation—Gal NAc‐T isoforms direct distinct subsets of the O‐glycoproteome
Article Snippet: .. Cleared acidified digests were loaded in equal amounts (peptide concentration) onto equilibrated SepPak C18 cartridges (Waters) followed by 3 × CV 0.1% TFA wash. Digests were labeled on‐column by adding 5 CV 30 mM NaBH3 CN and 0.2% formaldehyde in 50 mM sodium phosphate buffer pH 7.5 (L‐labeling), or 30 mM NaBH3 CN and 0.2% D‐formaldehyde in 50 mM sodium phosphate buffer pH 7.5 (M‐labeling). ..

Article Title: Targeting LIF-mediated paracrine interaction for pancreatic cancer therapy and monitoring
Article Snippet: .. Digested peptides from two biological replicate samples with stimulation and one control sample were de-salted using SepPak C18 cartridges in parallel and labeled using an on-column three-channel dimethyl labeling protocol . .. The labeled peptides were then combined at 1:1:1 ratio and phosphotyrosine (pTyr) peptides were immunoenriched by two rounds of immunoprecipitation with 4G10-conjugated agarose beads (Millipore) as described previously .

Article Title: De novo activating mutations drive clonal evolution and enhance clonal fitness in KMT2A-rearranged leukemia
Article Snippet: Peptides were differentially labeled with Tandem Mass Tag isobaric labeling (TMT 6-plex, Thermo Scientific) according to the manufacturer’s instructions with slight modifications. .. Following centrifugation, supernatants were collected and samples were desalted with SepPak C18 cartridges (Waters) by washing the columns with acetonitrile, conditioning and loading samples in 0.1% (v/v) trifluoroacetic acid, washing with 0.1% formic acid and eluting peptides with 80% (v/v) acetonitrile/0.1% (v/v) formic acid.

Article Title: Mammalian O-mannosylation of cadherins and plexins is independent of protein O-mannosyltransferases 1 and 2
Article Snippet: .. Trypsin digests were loaded onto equilibrated SepPak C18 cartridges (Waters) followed by three times CV of 0.1% TFA wash. Digests were labeled on the column by adding 5 ml of 30 m m NaBH3 CN and 0.2% formaldehyde (COH2 ) in 50 m m sodium phosphate buffer, pH 7.5 (Light, HEK293SC/PGNT1 ), or 30 m m NaBH3 CN and 0.2% deuterated formaldehyde (COD2 ) in 50 m m sodium phosphate buffer, pH 7.5 (Medium, HEK293SC/PGNT1/POMT1/POMT2 ). ..

Purification:

Article Title: Temperature has a major influence on cardiac natriuretic peptide in salmon
Article Snippet: The plasma samples were extracted with SepPak C18 cartridges (Waters, Milford, MA, USA) as described previously ( ) for use in sCP radioimmunoassay. .. Aliquots of the homogenates obtained during RNA purification (see above) were used to measure immunoreactive sCP (ir-sCP) in the tissue samples.

Article Title: HMGA1 regulates the Plasminogen activation system in the secretome of breast cancer cells
Article Snippet: .. iTRAQ labelling Each purified glycoprotein sample was dissolved with 500 μl of 0.1% formic acid, desalted on SepPak C18 Cartridges (Waters) and dried under vacuum. .. Each sample was suspended in 70 μl of dissolution buffer (iTRAQ Reagent Multi-Plex Kit, AB Sciex) and divided in two aliquots, each containing 25 μg of peptides. iTRAQ labelling was carried out essentially as previously described with a tag swapping strategy: Replicate 1 – I:114,116; NI:115,117/Replicate 2 – I:115,117; NI:116,114; Replicate 3 – I:116,114; NI:117,115.

Software:

Article Title: Quantifying exosome secretion from single cells reveals a modulatory role for GPCR signaling
Article Snippet: In brief, cells were lysed in denaturing lysis buffer supplemented with phosphatase inhibitors, lysates were digested overnight with sequence grade–modified trypsin (Promega), and 0.5-mg aliquots were desalted on SepPak-C18 cartridges (Waters). .. Protein identification was performed using MaxQuant software (1.5.2.8; ) with a FASTA file covering human canonical proteins and isoforms contained in the Swiss-Prot release of September 2015 (20,197 entries) using the following settings: The enzyme specificity was set to trypsin with a maximum tolerance of two missed cleavages.

Affinity Chromatography:

Article Title: A conserved major facilitator superfamily member orchestrates a subset of O-glycosylation to aid macrophage tissue invasion
Article Snippet: The cleared acidified digests were loaded onto equilibrated SepPak C18 cartridges (Waters) followed by 3 × CV 0.1% TFA wash. Digests were labeled on the column by adding 5 mL 30 mM NaBH3 CN and 0.2% formaldehyde (COH2 ) in 50 mM sodium phosphate buffer pH 7.5 (Light, mrva3102 ), or 30 mM NaBH3 CN and 0.2% deuterated formaldehyde (COD2 ) in 50 mM sodium phosphate buffer pH 7.5 (Medium, control). .. The eluates were mixed in a 1:1 ratio, concentrated by evaporation, and resuspended in Jacalin loading buffer (175 mM Tris-HCl, pH 7.4) Glycopeptides were separated from non-glycosylated peptides by Lectin Weak Affinity Chromatography (LWAC) using a 2.8 m column packed in-house with Jacalin-conjugated agarose beads.

Article Title: Quantifying exosome secretion from single cells reveals a modulatory role for GPCR signaling
Article Snippet: In brief, cells were lysed in denaturing lysis buffer supplemented with phosphatase inhibitors, lysates were digested overnight with sequence grade–modified trypsin (Promega), and 0.5-mg aliquots were desalted on SepPak-C18 cartridges (Waters). .. Then, phosphopeptides were captured through metal oxide affinity chromatography with in-house–prepared TiOx STAGE tips (packed with 2.5 mg 10-µm TiO2 beads; GL Sciences) in the presence of 300 mg/ml lactic acid to suppress background binding of acidic peptides, eluted with piperidine, desalted with in-house–prepared STAGE tips packed with SDB-XC (3M Empore), dried, and redissolved, and then 50% was injected into a nano–liquid chromatography–tandem mass spectrometry system (Ultimate 3000 nanoLC coupled on-line to a QExactive mass spectrometer; Thermo Fisher Scientific) operated as described previously by .

Article Title: Deconstruction of O‐glycosylation—Gal NAc‐T isoforms direct distinct subsets of the O‐glycoproteome
Article Snippet: Paragraph title: Dimethyl labeling and lectin weak affinity chromatography ... Cleared acidified digests were loaded in equal amounts (peptide concentration) onto equilibrated SepPak C18 cartridges (Waters) followed by 3 × CV 0.1% TFA wash. Digests were labeled on‐column by adding 5 CV 30 mM NaBH3 CN and 0.2% formaldehyde in 50 mM sodium phosphate buffer pH 7.5 (L‐labeling), or 30 mM NaBH3 CN and 0.2% D‐formaldehyde in 50 mM sodium phosphate buffer pH 7.5 (M‐labeling).

Sample Prep:

Article Title: De novo activating mutations drive clonal evolution and enhance clonal fitness in KMT2A-rearranged leukemia
Article Snippet: Paragraph title: Sample preparation for proteome analysis ... Following centrifugation, supernatants were collected and samples were desalted with SepPak C18 cartridges (Waters) by washing the columns with acetonitrile, conditioning and loading samples in 0.1% (v/v) trifluoroacetic acid, washing with 0.1% formic acid and eluting peptides with 80% (v/v) acetonitrile/0.1% (v/v) formic acid.

Evaporation:

Article Title: A conserved major facilitator superfamily member orchestrates a subset of O-glycosylation to aid macrophage tissue invasion
Article Snippet: The cleared acidified digests were loaded onto equilibrated SepPak C18 cartridges (Waters) followed by 3 × CV 0.1% TFA wash. Digests were labeled on the column by adding 5 mL 30 mM NaBH3 CN and 0.2% formaldehyde (COH2 ) in 50 mM sodium phosphate buffer pH 7.5 (Light, mrva3102 ), or 30 mM NaBH3 CN and 0.2% deuterated formaldehyde (COD2 ) in 50 mM sodium phosphate buffer pH 7.5 (Medium, control). .. The eluates were mixed in a 1:1 ratio, concentrated by evaporation, and resuspended in Jacalin loading buffer (175 mM Tris-HCl, pH 7.4) Glycopeptides were separated from non-glycosylated peptides by Lectin Weak Affinity Chromatography (LWAC) using a 2.8 m column packed in-house with Jacalin-conjugated agarose beads.

Article Title: Mammalian O-mannosylation of cadherins and plexins is independent of protein O-mannosyltransferases 1 and 2
Article Snippet: Trypsin digests were loaded onto equilibrated SepPak C18 cartridges (Waters) followed by three times CV of 0.1% TFA wash. Digests were labeled on the column by adding 5 ml of 30 m m NaBH3 CN and 0.2% formaldehyde (COH2 ) in 50 m m sodium phosphate buffer, pH 7.5 (Light, HEK293SC/PGNT1 ), or 30 m m NaBH3 CN and 0.2% deuterated formaldehyde (COD2 ) in 50 m m sodium phosphate buffer, pH 7.5 (Medium, HEK293SC/PGNT1/POMT1/POMT2 ). .. The eluates were mixed in a 1:1 ratio, concentrated by evaporation, and resuspended in ConA loading buffer (20 m m Tris-HCl, pH 7.4, 150 m m NaCl, 1 m m CaCl2 /MgCl2 /MnCl2 /ZnCl2 , 0.5 m urea).

Concentration Assay:

Article Title: Phosphoproteome Profiling Reveals Circadian Clock Regulation of Posttranslational Modifications in the Murine Hippocampus
Article Snippet: Each sample was diluted in fivefold volume of 50 mM NH4 HCO3 (pH 8.5) solution to reduce the urea concentration to < 2 M and digested overnight with TPCK-treated trypsin (Worthington, Lakewood, NJ, USA) at an enzyme-to-protein ratio of 1:25 (w/w). .. The remaining 1.1 mg of tryptic peptides were desalted by SepPak C18 cartridges (Waters, Mississauga, ON, Canada), dried, and SCX fractionated into four fractions (pH 4.0, 6.0, 8.0, and 12.0) prior to phosphoproteome enrichment.

Article Title: Deconstruction of O‐glycosylation—Gal NAc‐T isoforms direct distinct subsets of the O‐glycoproteome
Article Snippet: .. Cleared acidified digests were loaded in equal amounts (peptide concentration) onto equilibrated SepPak C18 cartridges (Waters) followed by 3 × CV 0.1% TFA wash. Digests were labeled on‐column by adding 5 CV 30 mM NaBH3 CN and 0.2% formaldehyde in 50 mM sodium phosphate buffer pH 7.5 (L‐labeling), or 30 mM NaBH3 CN and 0.2% D‐formaldehyde in 50 mM sodium phosphate buffer pH 7.5 (M‐labeling). ..

Liquid Chromatography with Mass Spectroscopy:

Article Title: Phosphoproteome Profiling Reveals Circadian Clock Regulation of Posttranslational Modifications in the Murine Hippocampus
Article Snippet: For proteomic analysis, 0.1 mg of resulting peptides were fractionated through an in-house constructed strong cation exchange (SCX) column with five pH fractions (pH 4.0, 6.0, 8.0, 10.0, and 12.0) followed by desalting with in-house C18 desalting cartridges and dried in a speed-vac prior to LC-MS analysis. .. The remaining 1.1 mg of tryptic peptides were desalted by SepPak C18 cartridges (Waters, Mississauga, ON, Canada), dried, and SCX fractionated into four fractions (pH 4.0, 6.0, 8.0, and 12.0) prior to phosphoproteome enrichment.

Article Title: Dynamic Impacts of the Inhibition of the Molecular Chaperone Hsp90 on the T-Cell Proteome Have Implications for Anti-Cancer Therapy
Article Snippet: The obtained peptide mixtures (200 µg total material) were desalted on SepPak C18 cartridges (Waters Corp., Milford, MA), dried, dissolved in 4M Urea with 0.1% Ampholytes pH 3–10 (GE Healthcare) and fractionated by off-gel focusing as described . .. The 24 fractions obtained were desalted on a microC18 96-well plate (Waters Corp., Milford, MA), dried and resuspended in 0.1% formic acid, 3% (v/v) acetonitrile for LC-MS analysis.

Lysis:

Article Title: Quantifying exosome secretion from single cells reveals a modulatory role for GPCR signaling
Article Snippet: .. In brief, cells were lysed in denaturing lysis buffer supplemented with phosphatase inhibitors, lysates were digested overnight with sequence grade–modified trypsin (Promega), and 0.5-mg aliquots were desalted on SepPak-C18 cartridges (Waters). .. Then, phosphopeptides were captured through metal oxide affinity chromatography with in-house–prepared TiOx STAGE tips (packed with 2.5 mg 10-µm TiO2 beads; GL Sciences) in the presence of 300 mg/ml lactic acid to suppress background binding of acidic peptides, eluted with piperidine, desalted with in-house–prepared STAGE tips packed with SDB-XC (3M Empore), dried, and redissolved, and then 50% was injected into a nano–liquid chromatography–tandem mass spectrometry system (Ultimate 3000 nanoLC coupled on-line to a QExactive mass spectrometer; Thermo Fisher Scientific) operated as described previously by .

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    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pre conditioned seppak c18 cartridge - by Bioz Stars, 2020-04
    86/100 stars
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